The Linker Histone H1.2 Is an Intermediate in the Apoptotic Response to Cytokine Deprivation in T-Effectors
Figure 2
H1.2 is an intermediate in the apoptotic cascade triggered by cytokine deprivation. (a)–(c) T-effectors were cultured in the indicated conditions (T0: freshly activated) and 6 hours later subject to subcellular fractionation. Immunoblots of nuclear and cytoplasmic fractions were probed for H1.2, HP1α, Bak, or Cox-IV. Data are representative of three independent trials. (d)-(e) T-effectors transduced using retrovirus expressing scrambled control or histone H1.2 shRNA, as described in Materials and Methods, were cultured with or without cytokine for 10 hours. Cells were scored for apoptotic nuclear damage using Hoechst-33342 (d) or mitochondrial transmembrane potential using TMRM (e) as described in Materials and Methods. Graphs show mean ± SD from 5 experiments in (d) and 2 experiments in (e), normalized to cultures with cytokine. Inset: Immunoblot for H1.2 protein and tubulin (parity in loading) in shRNA expressing T-effectors. *.
